The work in analytical biochemical laboratories and in clinical laboratories is often based on different tags or labels coupled on macromolecules under inspection. Typical labels used are different radioactive isotopes, enzymes, different fluorescent molecules and e.g. fluorescent chelates of rare earth metals. Detection of enzyme labels can be performed by utilizing its natural biochemical function, i.e. to alter the physical properties of molecules. In enzyme immunoassays colourless substances are catalysed by enzyme to colourful substances or non-fluorescent substances to fluorescent substances.
The colourful substances can be measured with absorption measurement, i.e. photometric measurement. In the absorption measurement the intensity of filtered and stabilized beam is first measured without any sample and then the sample inside one plate is measured. The absorbance i.e. the absorption values are then calculated.
The fluorescent substances can be measured with fluorescent measurement that is generally used for measuring quantities of fluorescent label substance in a sample. The most photoluminescence labels are based on molecular photoluminescence process. In this process optical radiation is absorbed by the ground state of a molecule. Due to the absorption of energy the quantum molecule rises into higher excited state. After the fast vibrational relaxation the molecule returns back to its ground state and the excess energy is released as an optical quantum. Due to losses in this process the average absorbed energies are higher than the average emitted energies.
A further measurement method is chemiluminescence measurement where emission of a substance is measured from a sample without excitation by illumination. Thus a photoluminometer suitable for photoluminescence measurements can also be used as a chemiluminometer.
Further, there is an analysing method called Amplified Luminescent Proximity Homogeneous Assay or AlphaScreen™. The function of the AlphaScreen™ method is based on the use of small beads that attach to the molecules under study. There are two types of beads that are coated with a material acting either as a donor or acceptor of singlet-state oxygen. The measurement starts, when the liquid sample is illuminated by light with a suitable wavelength e.g. 680 nm. After this, the material in the donor bead converts ambient oxygen into singlet-state oxygen. The single-state molecules have a short lifetime and they can reach only about a 200 nm distance by diffusion in the liquid. If the chemical reaction in question has taken place, both the donor and acceptor beads are bound to the same molecule and so they are sufficiently close to each other. In this case the singlet-state oxygen may reach the acceptor bead where a series of reactions is started. As the last phase of the reaction the coating material in the acceptor beads emits photons in the 500-700 nm range. If the chemical reaction has not taken place the singlet-state oxygen cannot reach the acceptor bead and the emission light is not detected. By measuring the intensity of light it is possible to conclude the efficiency of the chemical reaction.
An optical measurement instrument suitable for performing some or all of the measurements of the kind described above comprises typically at least one excitation light source for producing excitation beams to one or more samples to be measured at each time. Each excitation light source can be for example a flash lamp or a laser source. An optical path from an excitation light source to a sample may contain for example lenses, fibers, mirrors, dichroic mirrors, optical filters, monochromators and/or other optical elements. The optical measurement instrument further comprises at least one detector for detecting emission beams emitted by the samples to be measured at each time, and for producing detection signals responsive to the detected emission beams. The optical measurement instrument further comprises a processing device for producing a measurement result for each sample to be measured on the basis of the detection signal related to that sample. Each detector can be for example a photo-diode or a photo-multiplier tube. An optical path from the sample to the detector may contain for example lenses, fibers, mirrors, dichroic mirrors, optical filters, monochromators, and/or other optical elements. Each sample to be measured is contained by one of a plurality of sample wells that are built on e.g. a microtitration plate or another sample support element. In practical measurement circumstances temperatures of different samples may deviate from each other. The temperature deviation of different samples is a harmful phenomenon because the intensity of the emission radiation emitted by a sample depends on the temperature of the sample, and thus measurement results obtained from different samples are not commensurate with each other.
Publication U.S. Pat. No. 6,977,722 discloses an optical measurement instrument that includes one or more temperature sensors adapted to measure temperatures of a sample plate. Each temperature sensor may be a contact sensor, e.g. a temperature dependent resistor or a thermocouple, or a non-contact sensor such as an IR-sensor (infrared). The one or more temperature sensors is/are adapted to be able to measure the temperature of various locations on the sample plate, e.g. through the use of multiple sensors and/or by moving the sample plate relative to the sensors. The optical measurement instrument further comprises a computer adapted to receive the signal from the one or more temperature sensors, report the temperature to the user, and correct the measured signals to account for the effects of temperature. The accuracy of the temperature correction depends on temperature gradients on the sample plate and also on the selection of the locations on the sample plate from which the temperatures are measured.